Nucleic acid purification, such as the isolation of DNA or RNA, can be an important step in various biochemical and diagnostic procedures. RNA can be used in many applications, such as gene expression studies, molecular studies, and/or biochemical studies, for instance, in RNA interference, microinjection, infection, in vitro translation, and/or nuclease protection assay procedures. Transcription is the first step in gene expression, in which complementary and antiparallel RNA strands are synthesized from DNA template(s) using RNA polymerase. In vitro transcription (IVT) can provide a way to in vitro transcribe nucleotides with desired sequences or modifications, such as capping or radio labels, from DNA.
There are a variety of IVT kits designed to drive transcription, e.g., for T7, T3, or SP6 promoters. However, before continuing with downstream applications, it may be necessary to purify post-IVT samples to remove one or more contaminants, such as salts, proteins, enzymes, oligonucleotides, and the like. The presence of such contaminating materials can impede or prevent many downstream processes. Thus, it can be important to effectively isolate nucleic acids from the post-IVT mixture to ensure a desired end-use functionality.
The selected method for purifying nucleic acids can, in some instances, impact various properties of the isolated product, including yield, quality, and/or purity of the nucleic acid sample. While many approaches have been developed for nucleic acid purification, these methods may have one or more drawbacks including, for example, high cost, high complexity, slow speed, low yield, low purity, contamination, toxicity, and/or inefficiency. RNA of relatively high purity can be isolated from IVT mixtures using traditional precipitation procedures, such as phenol/chloroform extraction; however, such methods can be time-consuming and complex.
Solid phase-based methods, such as methods utilizing magnetic beads, spin columns, and/or filtration systems, have been presented as an alternative solution. Among these methods, magnetic particle-based purification systems may have various advantages, such as enhanced simplicity, due to the lack of centrifugation and/or vacuum steps. However, methods employing magnetic particles can still suffer from various disadvantages, such as slow speed, high complexity, and/or poor overall yield.
Accordingly, it would be advantageous to provide magnetic particle-based methods and kits for nucleic acid purification that may be faster, less complex, less expensive, and/or improved in terms of product purity and/or yield. The resulting purified nucleic acids can be used in a wide variety of gene expression, molecular, and/or biochemical applications.